Producing Peptostreptococcus Magnus Protein L and binding domain using E. Coli

Protein L is an antibody-binding protein that interacts with the light chains. There are multiple domains within the protein and there could be a possibility to increase the binding capabilities. The area of protein L that binds to the antibody comprises several domains, the focus is the B1 domain. The protein cannot be produced on its own, it must be produced using a host organism. The host organism is BL21 and this was picked because it has the opportunity to allow proteins to grow due to the absence of proteases that would usually break down proteins. The next factor taken into consideration was the plasmid that would contain the gene of interest, the plasmid chosen was pET 28a, and the plasmid was chosen based on its antibiotic resistance to kanamycin, which would allow for the selection process to isolate the plasmids possibly containing the gene of interest. Lastly, the cut enzymes were chosen because of their location in contrast to the 6x His Tag. This project will be expanded into future processes such as the mass production of the protein and purifying the protein even further.